| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
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| Targets |
Fluorescent Dye
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|---|---|
| ln Vitro |
I. Reagent Preparation
1. Protein Preparation Requirements: • Optimal concentration: 2 mg/mL (range: 2-10 mg/mL) • pH adjustment: Maintain 8.5±0.5 using 1M sodium bicarbonate if pH<8.0 • Buffer compatibility: Must be free of primary amines (Tris/glycine) and ammonium ions 2. Dye Solution Preparation: • Dissolve VF dye in anhydrous DMSO to 10 mg/mL • Mix thoroughly using vortex or glass rod • Storage: Aliquot and protect from light at -20°C/-80°C 3. Dye Volume Calculation (Example for IgG): Given: 500μL of 2mg/mL IgG (MW=150kDa), 1mg VF488 (MW=834) Calculation steps: 1. IgG mmol = (2mg/mL×0.5mL)/150,000 = 6.7×10⁻⁶ mmol 2. VF488 mmol = 6.7×10⁻⁶ ×10 = 6.7×10⁻⁵ mmol 3. VF488 μL = (6.7×10⁻⁵×834)/0.01 = 5.6μL II. Labeling Procedure 1. Conjugation Reaction: • Slowly add calculated VF dye to protein solution • Gentle mixing followed by brief centrifugation • Incubate at RT for 60min with periodic inversion (every 10-15min) 2. Purification (Sephadex G-25): • Prepare column per manufacturer's instructions • Load reaction mixture onto column • Elute with PBS (pH7.2-7.4) • Collect conjugated protein fractions Storage: -20°C protected from light Key Considerations: 1. Light/moisture sensitivity: Prepare dye fresh for each use 2. Compatible preservatives: ≤3mM NaN₃ or ≤0.02mM thimerosal 3. Incompatible components: Glycerol (20-50%), amine-containing buffers 4. Safety: For research use only with proper PPE |
| References | |
| Additional Infomation |
N-hydroxysuccinimde (NHS) ester-mediated derivitization involves the reaction of this amine-reactive group with the primary amines of a protein or a biomolecule. Using NHS chemistry allows one to conjugate various fluorescent probes, biotin, and cross-linkers to primary amines. For example, we use NHS ester chemistry to fluorescently label the amino terminus of a protein with the dye, 5-(and-6)-carboxyfluorescein, succinimidyl ester (5(6)-FAM, SE). [1]
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| Appearance |
Purple to purplish red solid powder
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|---|---|
| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
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